Mechanisms of Norovirus Pathogenesis and Replication to Develop Therapeutics

诺如病毒致病机制和复制治疗药物的研究进展 项目摘要 诺如病毒是美国胃肠炎的最常见原因，它们导致显著的 给人民带来经济和健康负担。这是有效疗法取得进展的一个重大障碍， 控制人类诺沃克病毒疾病的抗病毒药物和疫苗是以前缺乏细胞培养系统。焦点 该计划项目的基础是建立在以前的基础科学和翻译发现和发现的基础上，以允许 我们对这些病毒的生物学和发病机制的了解不断取得进展，从而改善了 可以制定控制策略。我们最近开发了一种成功的复制系统，它发现了菌株- 复制的特殊要求，并允许中和分析和抗病毒药物进行测试。我们将追查关键 关于病毒致病机制和细胞反应的问题，关于病毒与病毒相互作用的结构-功能研究 病毒蛋白和宿主成分，发现限制广泛传播的关键因素并测试抗病毒药物 以及在三个不同的项目中中和抗体。在项目1中，我们将进行旨在提供 我们对人类诺如病毒如何致病以及哪些表位有基本的分子理解 可被与病毒中和、病毒清除和保护有关的人类血清识别 豁免权。此外，我们将在最近成功的基础上，继续开发用于治疗和预防的抗病毒药物 可应用于高危人群的策略。在项目2中，我们将继续完善种植制度， 专注于识别细胞受体(S)并理解为什么所有的户新病毒毒株都不能在复制中生长 系统。我们还将评估诺沃克病毒复制调节细胞固有的分子机制。 以及这些细胞反应是否调控病毒的复制和传播。在项目3中，我们将 确定调节病毒复制和病毒宿主的关键蛋白新功能的结构基础 相互作用，为抗病毒药物的开发提供合理的框架。这些项目将得到支持 三核之差。核心A(行政核心)将提供集中的行政和财务管理 支持并将协调方案活动。核心B(显微镜和肠道核心)将提供 为每个项目提供与电子显微镜和荧光显微镜相关的专业知识和服务。这个核心将 还为所有项目维护和提供肠样体，包括转基因培养物和生物传感器线路。 核心C(蛋白质和小分子化学核心)将为所有项目提供获得纯化蛋白质的途径 和病毒样颗粒，以及促进所需的定点突变活动。此外，核心将 合成小分子，用于每个项目中的蛋白酶和聚合酶抑制研究。该计划 该项目汇集了一组具有不同技能和才华的高度协作的调查人员，以及 在一起工作的悠久历史。与上一个资助期一样，每个项目之间的互动 每个核心都将具有广泛性和协同性，从而加强每个项目的活动 如果这些项目是独立进行的，可以取得什么成就，这一点是相当大的。

Mechanisms of Norovirus Pathogenesis and Replication to Develop Therapeutics Project Summary Noroviruses are the most common cause of gastroenteritis in the United States, and they cause a significant economic and health burden to the population. A significant barrier to progress in making effective therapeutics, antivirals and vaccines to control human norovirus illness was the previous lack of a cell culture system. The focus of this program project is to build upon previous basic science and translational findings and discoveries to allow continued progress in our understanding of the biology and pathogenesis of these viruses so that improved control strategies can be developed. We recently developed a successful replication system that discovered strain- specific requirements for replication and allows neutralization assays and antivirals to be tested. We will pursue key questions on virus pathogenesis and cellular responses, structure-function studies on the interactions between virus proteins and host components, discover key factors that restrict extensive propagation and test antivirals and neutralizing antibodies in three separate projects. In Project 1 we will perform studies designed to provide us with a fundamental molecular understanding of how human noroviruses cause disease, and of what epitopes are recognized by human sera that are associated with virus neutralization, virus clearance, and protective immunity. In addition, we will build on recent success and continue to develop antivirals for treatment and prevention strategies that can be applied to at risk populations. In Project 2 we will continue to improve the cultivation system, focusing on identifying the cell receptor(s) and understanding why all HuNoV strains do not grow in the replication system. We will also evaluate molecular mechanisms by which norovirus replication regulates cellular innate responses and whether these cellular responses regulate viral replication and spread. In Project 3 we will determine the structural basis for novel functions of key proteins that regulate viral replication and virus-host interactions to provide a rational framework for the development of antivirals. These projects will be supported by three cores. Core A (Administrative Core) will provide centralized administrative and fiscal management support and will coordinate programmatic activities. Core B (the Microscopy and Enteroid Core) will provide expertise and services to each project related to electron microscopy and fluorescent microscopy. This core will also maintain and provide enteroids, including genetically-modified cultures and biosensor lines to all projects. Core C (the Protein and Small Molecule Chemistry Core) will provide all projects with access to purified proteins and virus-like particles as well as facilitating site-directed mutagenesis activities needed. In addition, the Core will synthesize small molecules to be used for protease and polymerase inhibition studies in each project. The program project brings together a highly collaborative group of investigators with diverse skills and talents, and an substantial history of working together. As in the previous funding period, the interactions among each project and each of the cores will be extensive and synergistic such that the activities of each project will be enhanced considerably over what could be accomplished if the projects were pursued independently.